Study On Extractions,Component Analyses And Activities Of The Oil From Rana Dybowskii Eggs

In this study,ovum oil of Rana chensinensis（OORC）was extracted from dried Rana dybowskii eggs as raw materials,in which the extraction rate of OORC as the observation index by comparing a few of process parameters of reflux extraction,supercritical CO₂extraction and subcritical extraction to determine the optimized method.The OORC composition was assayed by gas chromatography-mass spectrometry（GC-MS）,and the activity of the OORC was analyzed and verified by the network pharmacology.The main results of this paper were as follows:1.The OORC was extracted by reflux extraction,supercritical CO₂extraction and subcritical extraction,and three different extraction methods were optimized.Finally,the extraction rate of OORC（34.96%）obtained by supercritical CO₂extraction was the highest and the best quality.The optimized extraction parameters were temperature 60°C,length 150 min,pressure 36 MPa,CO₂flow rate 10 L/h.2.The composition of the OORC was assayed by GC-MS and 34 kinds of fatty acids were determined in the oil,in which the contents of unsaturated fatty acid and saturated fatty acid are 68.05%and 31.95%,respectively.3.Based on the network pharmacology,the compositions of the oil with relative content more than 1%were analyzed by using STP database to obtain the corresponding targets of the OORC;the key target genes were analyzed for gene ontology（GO）and Kyoto Encyclopedia of Genes and Genomes（KEGG）using Clue GO plug-in and Metascape database;the network of"component-target-pathway"of the OORC was constructed using Cytoscape software,and the results implied that it had potential advantages in anti anxiety,anti-oxidation and lipid lowering.4.No death was observed in the acute toxicity test of rats with Intragastric dosage 10.0g/（kg?bw）,and LD₅₀was zero.With Maximum Tolerable Dose（MTD）test under the acceptable maximum dosage of the drug was upto 80.0 g/（kg.bw）in 24 hours,and no signs of poisoning were found in rats,indicating that edible OORC was safe and reliable.5.With animal anxiety models,in two behavioral tests,Elevated Plus Maze（EPM）and Light and dark box（LAB）,controling the study and control groups,the frequency/persistence of the mice entering the tracks in the study group were significantly higher/longer than one in the control group,individually.the OORC having a strong anti anxiety effect.6.With the antioxidant activity tests,the results showed that the half inhibitory concentration(IC₅₀)values of radical scavenging,hydroxyl radical scavenging and superoxide anion scavenging against 1,1-diphenyl-2-trinitrophenylhydrazine（DPPH）were0.897,1.392 and 1.687,individually,the OORC having very strong antioxidant activity in vitro.7.The hyperlipidemia rat model successfully established with high-fat diet for 28days,in which the rats were given different substances by gavage.With data comparison and analyses,it was found that the OORC had significant effects on triglyceride,LDL cholesterol and total cholesterol in hyperlipidemia rats.